作物学报 ›› 2009, Vol. 35 ›› Issue (7): 1202-1208.doi: 10.3724/SP.J.1006.2009.01202
牛义1,王志敏1,高启国1,宋明1,王小佳1,*,朱利泉2,*
NIU Yi1,WANG Zhi-Min1,GAO Qi-Guo1,SONG Ming1,WANG Xiao-Jia1*,ZHU Li-Quan2*
摘要:
在甘蓝自交不亲和信号传导中ARC1和上游因子SRK之间可能存在相互作用。为进一步证实该相互作用,以甘蓝E1为材料,采用RT-PCR技术扩增ARC1的编码序列, 构建ARC1原核表达质粒pET43.1a-ARC1,转化宿主菌大肠杆菌BL21,通过SDS-PAGE检测该蛋白的表达。利用免疫共沉淀原理及pET43.1a-ARC1融合蛋白序列中的6×His标签与Ni+结合的特点建立了体外检测蛋白质相互作用的新方法, 并用该方法对ARC1与SRK的相互作用进行了检测。结果表明,在体外ARC1能与SRK相互作用并形成复合体,这为深入分析ARC1与SRK相互作用机理以及探讨ARC1与下游传导元件的相互作用提供了理论和技术基础。
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